Electron Microscopy Grants and Publications

Descriptions for Grant Writers 

Any use of the resources at the Fred Hutchinson Cancer Center Electron Microscopy shared resource requires acknowledgement of the core in publication as follows: “Electron microscopy data were generated using the Fred Hutchinson Cancer Center Electron Microscopy shared resource. The EMSR is supported in part by the Cancer Center Support Grant P30 CA015704-40.” Descriptions of the overall Fred Hutch Shared Resources program are available on the main Shared Resources grant information page.

Examples of publications made possible by the Electron Microscopy shared resource staff are listed below.


Citations for CCSG  |  Grant Descriptions  |  Publications

Citations for CCSG-Support Research

All publications, press releases, or other documents that cite results from CCSG-supported research must include acknowledgement of the grant and maintain compliance with NIH Public Access Policy. All manuscripts accepted for publication must be submitted to PubMed Central and be assigned a PMCID. Additionally, please reference the Research Resource Identifier (RRID). RRIDs are assigned to cores to help researchers cite key resources in the biomedical literature to improve transparency of research methods.

“This research was supported by the Electron Microscopy Shared Resource, RRID:SCR_022611, of the Fred Hutch/University of Washington/Seattle Children’s Cancer Consortium (P30 CA015704).”

Electron Microscopy Grant Descriptions

Short Grant Description

The Electron Microscopy shared resource provides labs with training and access to two high-resolution transmission electron microscopes, a scanning electron microscope, and the necessary sample-preparation tools. This shared resource offers negative-stain, embedding, sectioning and vitrification tools, a Talos L120C TEM, a Glacios 200kV TEM, four A6000 GPUs, eight NVIDIA RTX3090 GPUs, and automated data-collection software. Staff are trained in sample preparation and high-resolution image acquisition for TEM and SEM and the entire workflow of structure determination by cryo-EM. Training and full service sample preparation and imaging are offered to both internal and external users.

Long Grant Description 

The Electron Microscopy shared resource provides users with the support and resources needed to determine structures of biological specimens to further their scientific pursuits. The electron microscopy suite at Fred Hutchinson Cancer Center is newly renovated to accommodate 200 kV Glacios and 120 kV Talos transmission microscopes and a 30kV JEOL JSM 6610LV scanning electron microscope. These are available for assisted and independent use after training provided by staff members in the EM shared resource. In the same approximately 550 square feet of space, all tools and instruments needed for tissue preparation, cellular preparation, negative stain and sample vitrification are provided. Shared resource staff maintain the microscopes and sample-preparation equipment in a fully functional state with alignments verified bimonthly. They also train new and experienced users in the operation of the hardware and software provided by the core, detailed below.

The Talos microscope is equipped with a Ceta 16M CMOS camera for high-speed and high-resolution 4k x 4k imaging in low-dose mode. This microscope is used for negative-stain data collection and cryo-EM screening with our room-temperature and cryogenic Elsa holder. The Glacios microscope is equipped with a K3 camera, the newest generation of direct electron detector from Gatan. This camera has a large sensor that can operate at 1500 frames per second, allowing for exceptionally sensitive and fast data collection.

The shared resource is also supported by Fred Hutch Shared Resources and Center IT to provide 10 GbE data-transfer connections, enabling on-the-fly drift correction and initial live processing during imaging sessions on the Glacios. These data are retained for one month on the shared resource’s storage server, and users are provided with options for transferring data off this server internally and externally. Both microscopes have SerialEM and Leginon installed and calibrated for use in automated data collection, which enable the collection of dozens of micrographs per hour and tomographic data collection. Image processing is supported by individual labs, but staff provide office hours and individual consultations on adjusting data-processing parameters for optimal results.

Publications by Electron Microscopy

Our team in the Electron Microscopy shared resource is proud to have contributed to major scientific advances. Following is a sample of the high-impact publications our work has made possible:

More Publications

  • Shahbazi R, Sghia-Hughes G, Reid JL, Kubek S, Haworth KG, Humbert O, Kiem HP, Adair JE. Targeted homology-directed repair in blood stem and progenitor cells with CRISPR nanoformulations. Nat Mater. 2019 Oct;18(10):1124-1132. doi: 10.1038/s41563-019-0385-5. Epub 2019 May 27. PMID: 31133730; PMCID: PMC6754292.
  • Taylor JA, Bratton BP, Sichel SR, Blair KM, Jacobs HM, DeMeester KE, Kuru E, Gray J, Biboy J, VanNieuwenhze MS, Vollmer W, Grimes CL, Shaevitz JW, Salama NR. Distinct cytoskeletal proteins define zones of enhanced cell wall synthesis in Helicobacter pylori. Elife. 2020 Jan 9;9:e52482. doi: 10.7554/eLife.52482. PMID: 31916938; PMCID: PMC7012605.
  • Correnti CE, Hallinan JP, Doyle LA, Ruff RO, Jaeger-Ruckstuhl CA, Xu Y, Shen BW, Qu A, Polkinghorn C, Friend DJ, Bandaranayake AD, Riddell SR, Kaiser BK, Stoddard BL, Bradley P. Engineering and functionalization of large circular tandem repeat protein nanoparticles. Nat Struct Mol Biol. 2020 Apr;27(4):342-350. doi: 10.1038/s41594-020-0397-5. Epub 2020 Mar 23. PMID: 32203491; PMCID: PMC7336869.
  • Verboon JM, Nakamura M, Davidson KA, Decker JR, Nandakumar V, Parkhurst SM. *Drosophila* Wash and the Wash regulatory complex function in nuclear envelope budding. J Cell Sci. 2020 Jul 8;133(13):jcs243576. doi: 10.1242/jcs.243576. PMID: 32503943; PMCID: PMC7358131.
  • Bruchez A, Sha K, Johnson J, Chen L, Stefani C, McConnell H, Gaucherand L, Prins R, Matreyek KA, Hume AJ, Mühlberger E, Schmidt EV, Olinger GG, Stuart LM, Lacy-Hulbert A. MHC class II transactivator CIITA induces cell resistance to Ebola virus and SARS-like coronaviruses. Science. 2020 Oct 9;370(6513):241-247. doi: 10.1126/science.abb3753. Epub 2020 Aug 27. PMID: 32855215; PMCID: PMC7665841.
  • Shen BW, Quispe JD, Luyten Y, McGough BE, Morgan RD, Stoddard BL. Coordination of phage genome degradation versus host genome protection by a bifunctional restriction-modification enzyme visualized by CryoEM. Structure. 2021 Jun 3;29(6):521-530.e5. doi: 10.1016/j.str.2021.03.012. Epub 2021 Apr 6. PMID: 33826880; PMCID: PMC8178248.
  • Mosquera JV, Bacher MC, Priess JR. Nuclear lipid droplets and nuclear damage in Caenorhabditis elegans. PLoS Genet. 2021 Jun 16;17(6):e1009602. doi: 10.1371/journal.pgen.1009602. PMID: 34133414; PMCID: PMC8208577.
  • Al Qureshah F, Sagadiev S, Thouvenel CD, Liu S, Hua Z, Hou B, Acharya M, James RG, Rawlings DJ. Activated PI3Kδ signals compromise plasma cell survival via limiting autophagy and increasing ER stress. J Exp Med. 2021 Dec 6;218(12):e20211035. doi: 10.1084/jem.20211035. Epub 2021 Sep 29. PMID: 34586341; PMCID: PMC8485856.
  • Hallinan JP, Doyle LA, Shen BW, Gewe MM, Takushi B, Kennedy MA, Friend D, Roberts JM, Bradley P, Stoddard BL. Design of functionalised circular tandem repeat proteins with longer repeat topologies and enhanced subunit contact surfaces. Commun Biol. 2021 Oct 29;4(1):1240. doi: 10.1038/s42003-021-02766-y. PMID: 34716407; PMCID: PMC8556268.
  • Nagao RJ, Marcu R, Shin YJ, Lih D, Xue J, Arang N, Wei L, Akilesh S, Kaushansky A, Himmelfarb J, Zheng Y. Cyclosporine Induces Fenestra-Associated Injury in Human Renal Microvessels *In Vitro*. ACS Biomater Sci Eng. 2022 Jan 10;8(1):196-207. doi: 10.1021/acsbiomaterials.1c00986. Epub 2021 Dec 19. PMID: 34927415.
  • Hao S, Inamdar VV, Sigmund EC, Zhang F, Stephan SB, Watson C, Weaver SJ, Nielsen UB, Stephan MT. BiTE secretion from in situ-programmed myeloid cells results in tumor-retained pharmacology. J Control Release. 2022 Feb;342:14-25. doi: 10.1016/j.jconrel.2021.12.029. Epub 2021 Dec 23. PMID: 34953983; PMCID: PMC8840964.
  • El-Kharrag R, Berckmueller KE, Madhu R, Cui M, Campoy G, Mack HM, Wolf CB, Perez AM, Humbert O, Kiem HP, Radtke S. Efficient polymer nanoparticle-mediated delivery of gene editing reagents into human hematopoietic stem and progenitor cells. Mol Ther. 2022 Jun 1;30(6):2186-2198. doi: 10.1016/j.ymthe.2022.02.026. Epub 2022 Feb 28. PMID: 35240320; PMCID: PMC9171380.
  • Stolla MC, Reilly A, Bergantinos R, Stewart S, Thom N, Clough CA, Wellington RC, Stolitenko R, Abkowitz JL, Doulatov S. ATG4A regulates human erythroid maturation and mitochondrial clearance. Blood Adv. 2022 Jun 28;6(12):3579-3589. doi: 10.1182/bloodadvances.2021005910. PMID: 35443024; PMCID: PMC9631553.
  • Piliponsky AM, Sharma K, Quach P, Brokaw A, Nguyen S, Orvis A, Saha SS, Samanas NB, Seepersaud R, Tang YP, Mackey E, Bhise G, Gendrin C, Furuta A, Seo AJ, Guga E, Miralda I, Coleman M, Sweeney EL, Bäuml CA, Imhof D, Snyder JM, Moeser AJ, Rajagopal L. Mast cell-derived factor XIIIA contributes to sexual dimorphic defense against group B streptococcal infections. J Clin Invest. 2022 Oct 17;132(20):e157999. doi: 10.1172/JCI157999. PMID: 36006736; PMCID: PMC9566924.
  • Sichel SR, Bratton BP, Salama NR. Distinct regions of *H. pylori'*s bactofilin CcmA regulate protein-protein interactions to control helical cell shape. Elife. 2022 Sep 8;11:e80111. doi: 10.7554/eLife.80111. PMID: 36073778; PMCID: PMC9507126.
  • Wicky BIM, Milles LF, Courbet A, Ragotte RJ, Dauparas J, Kinfu E, Tipps S, Kibler RD, Baek M, DiMaio F, Li X, Carter L, Kang A, Nguyen H, Bera AK, Baker D. Hallucinating symmetric protein assemblies. Science. 2022 Oct 7;378(6615):56-61. doi: 10.1126/science.add1964. Epub 2022 Sep 15. PMID: 36108048; PMCID: PMC9724707.
  • Bean EM, Morella NM, Dey N. Complete Genome Sequences of Two Bacteroides uniformis Bacteriophages. Microbiol Resour Announc. 2022 Oct 20;11(10):e0061022. doi: 10.1128/mra.00610-22. Epub 2022 Sep 19. PMID: 36121235; PMCID: PMC9584325.
  • Cheng RY, Hung KL, Zhang T, Stoffers CM, Ott AR, Suchland ER, Camp ND, Khan IF, Singh S, Yang YJ, Rawlings DJ, James RG. Ex vivo engineered human plasma cells exhibit robust protein secretion and long-term engraftment in vivo. Nat Commun. 2022 Oct 16;13(1):6110. doi: 10.1038/s41467-022-33787-8. PMID: 36245034; PMCID: PMC9573882.